The diversity in Tibetan kefir grains and dynamics of the microbial community during the fermentation of Tibetan kefir by PCR-DGGE fingerprinting technique were studied.The results showed that bacterial community of Tibetan kefir grains was more complex than that of yeast.The bacteria communities between different Tibetan kefir grains showed 78%～84% similarity, and yeast 80%～92%.Bands B, E and N of DGGE profiles of the microbial community during fermentation of Tibetan kefir were present throughout the fermentation.Analysis of sequence date showed the majority of the DGGE bands of bacteria community corresponded to LAB, and the most intense band (band E) was completely homology to Lactococcus lactis.

Objective To investigate the intensity of HBsAg and HBcAg expression in liver tissue of patients with chronic hepatitis B virus (HBV) infection and its clinical significance.Methods A total of 994 HBV infected patients underwent liver biopsy and histopathological examination.The expression of HBsAg and HBcAg in liver tissue was detected by histoimmunochemistry.Patients were divided into HBeAg (+)/HBVDNA(+), HBeAg(-)/HBV DNA(+) and HBeAg(-)/HBV DNA(-) groups according to HBeAg and HBV DNA levels;patients were divided into ＜2 × normal (ULN) group, 2-＜5 × ULN groupand ≥5 × ULN group according to the alanine aminotransferase (ALT) levels.The histologic activity (A), fibrosis (F), the expression of HBsAg and HBcAg in liver tissue and their correlations with clinical features were analyzed.Logistic regression analysis was used to study the factors affecting the expression of HBsAg and HBcAg in liver tissue.Results Among 994 HBV infected patients, 941 cases (94.67％) were intrahepatic HBsAg positive and 553 cases (55.63％) were intrahepatic HBcAg positive;403 cases (40.85％) were ≥A2 in histologic activity and 371 cases (36.09％) were ≥F2 in fibrosis.The degree of A and F was the highest in HBeAg (-) / HBV DNA (+) group, followed by HBeAg (-) / HBV DNA (-) group, and was the lowest in HBeAg (+) / HBV DNA (+) group.The intensity of intrahepatic HBsAg expression was significantly different among three groups (x2 =6.299, r =-0.760, P ＜ 0.05), however, the difference was not showed in pairwise comparisons.The difference of intrahepatic HBcAg intensity among three groups was statistically significant (x2 =282.995, r =-0.645, P ＜ 0.01), the intensity was the highest in HBeAg (+) / HBV DNA (+) group and the lowest in HBeAg (-) / HBV DNA (-) group.The constituent ratio of HBeAg positive and HBV DNA level were higher and the average age was lower in intrahepatic HBsAg positive group than those in HBsAg negative group.The constituent ratio of positive HBeAg, the levels of ALT, AST, PLT and HBV DNA were higher and the average age, the average FIB-4 level were lower in intrahepatic HBcAg positive group than those in HBcAg negative group.The HBV DNA level was an independent risk factor for intrahepatic HBsAg intensity, and the HBeAg positive and HBV DNA level were independent risk factors for intrahepatic HBcAg intensity.There were no significant differences in A and F among different groups of intrahepatic HBsAg intensity (x2 =1.943 and 2.630, both P ＞ 0.05).There was significant difference in F among different groups of intrahepatic HBcAg intensity (x2 =12.352, P ＜ 0.01), but not in A.The degree of F was the highest in intrahepatic HBcAg negative group.There was significant difference in intrahepatic HBcAg intensity among different groups of ALT level (x2 =16.349, P ＜ 0.01), but not in intrahepatic HBsAg intensity.The intrahepatic HBcAg intensity in ALT ＜ 2 × ULN group was lower than that in other two groups.Conclusions Most of patients with chronic HBV infection are intrahepatic HBsAg positive and more than half of them are intrahepatic HBcAg positive.The intrahepatic HBsAg intensity is not associated with A and F, but correlates with HBV DNA level.The intrahepatic HBcAg intensity is not associated with A, but it is negatively correlated with F and positively correlated with positive HBeAg expression, HBV DNA level and ALT level.

Objective To investigate the effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems of school-aged children in Shanghai. Methods A total of 4 108 school-aged children from 10 primary schools of Shanghai were enrolled by multi-stage cluster sampling and surveyed by questionnaires. The information of television-watching and computer-using, family and personal condition was investigated by self-prepared questionnaire, and the Chinese version of Children's Sleep Habits Questionnaire was employed to survey the sleep behaviors of children. The effects of television-watching and computer-using on sleep/wake patterns, sleep duration and sleep problems were analyzed by multiple linear regression analysis and Logistic regression analysis. Results The percentage of children who watched television≥2 h per day was 4.1% during weekdays, and that came to 49.2% during weekends. In terms of frequency of computer-using, most children reported "rarely" (88.2%, 0-1 time/week), followed by "often" (11.0%, 2-4 times/ week) and "usually" (0.8%, 5-7 times/week). With the age increase, the percentages of children who watched television≥2 h per day and those who "often" used computer gradually increased. It was revealed by multiple linear regression analysis and Logistic regression analysis that television-watching and computer-using were not only positively correlated with later bedtime, later wake time and shorter sleep duration but also significantly associated with sleep problems such as bedtime resistance, sleep onset delay, sleep duration disorder, sleep anxiety and parasomnia. Conclusion Television-watching and computer-using exert influences on sleep behaviors of sleep/wake patterns, sleep duration and sleep problems. Concerns about the potential negative effects of television-watching and computer-using on sleep behaviors may help to promote healthy sleep patterns and improve sleep quality.

Objective To evaluate the clinical significance of the diameter of the esophageal hiatus on multislice spiral CT(MSCT)and to present the MSCT manifestations of esophageal hiatus hernia (EHH).Methods(1)The distance between diaphragmatic crura(DDC),which indicated the diameter of esophageal hiatus,was measured in 140 normal adult patients on their thoracic and/or abdomenal CT images.(2)The DDC of 56 patients with EHH diagnosed by barium examination was measured on MSCT, and the MSCT findings were analyzed retrospectively.Results(1)The DDC of 140 normal adult cases were(13.44?4.41)mm on average and increased with age.The mean DDCs of patients under the age of 59 year-old(80 cases)and over 60-year-old(60 cases)were 11.03?2.10 mm and 16.67?4.64 mm respective]y,there was a significant difference(t=8.762,P

Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual funetion.However,the color vision system of animals which encounter uncertainty of spectral signals should be plastic.Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear.Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light.Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed tO the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively.The flat-mounted retinal sample was prepared and divided into dorsal zone,ventral zone and mixed zone anatomically according to the distribution of difierent light-sensitive cone.The changes in density of cone cells sensitivited to different colored light were detected by single-1abel or double-label immunocytochemistry.The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively.Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group,purple light group and white light group (F=234.28,P<0.01).Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group wag evidently lower(q=389.68,P<0.01；q=67.11,P<0.01).No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group,purple light group and white light group(F=3.14,P>0.05).The density of coexpression of the mixed cone cells was increased in green light group(q=157.55,P<0.01)but decreased in purple light roup (q=254.85,P<0.01)in comparison with white light group.The expression levels of green-opsin and green-opsin mRNA in green light group was significantly elevated(q=184.45,P<0.01；q=4.71,P<0.05),but those of purple light group were evidently declined(q=5.87,P<0.05；q=346.66,P<0.01)in comparison with white light group.There was no statistically significant differences were found in the expression of purple-opsin and its mRNA among all the groups(F=1.24,P>0.05；F=3.27,P>0.05).Conclusion After the exposure of long-time monochromatic light illumination,monochromatic cones density and its opsin in guinea pig occur the corresponding alteration to gain good spatial vision as a compensatory reaction.These outcomes imply that there is some plasticity during the development of color vision.The increase of green-sensitive cones might be from the differentiation of coexpression cones in transition region.

Objective To discuss the effects of acupuncture therapy on resting state functional magnetic resonance imaging (fMRI) in patients with ischemic stroke; To explore the central mechanism of acupuncture therapy.Methods Ten is chemic stroke patients with hemiplegia limbs and ten matched control subjects were included. Treatment group was given anti-platelet aggregation, anticoagulation, and nutrition brain cell therapy. At the same time, acupuncture therapy was given, taking Jianyu, Quchi, Shousanli, Waiguan, Hegu, Zusanli, Fenglong, Yanglingquan, Sanyinjiao, and Taichong, with even reinforcing-reducing method, and retaining the needle 30 min, manipulating needle every 10 min, once a day, 5 times a week, needle stopping for 2 d, for continuous 30 d. The health group did not undergo any treatment. BOLD-fMRI imaging technique was used, and based on RSET1.8 software, the parameters of brain activity before and after treatment were observed by amplitude of low frequency fluctuation (ALFF) method. The Results were compared with healthy group. Results Before treatment, the ALFF value of left caudate nucleus, putamen and globus pallidus in the treatment group was lower than that in the health group, and the ALFF value of bilateral precentral and postcentral gyrus, left grus frontalis superior, gyrus frontalis medius, gyrus temporalis medius and precuneus was higher than the health group (P<0.05). After treatment, the ALFF value in left caudate nucleus, putamen and globus pallidus in the treatment group was higher than that before treatment, and the ALFF value in left gyrus frontalis superior, gyrus frontalis medius and precuneus decreased than before group (P<0.05).Conclusion In the resting state of the ischemic stroke patients with hemiplegia limbs, the principal injured brain regions are caudate nucleus, putamen and globus pallidus. Acupuncture has a regulatory role for these brain areas, presumably may be the central mechanism of efficacy.

AlM:To evaluate the clinical efficacy of intravitreal injection of ranibizumab combined with macular grid photocoagulation for diabetic macular edema ( DME) .METHODS:Totally 60 eyes ( 60 patients ) with DME were randomly divided into 2 groups: 30 eyes of simple injection group underwent intravitreal injection of ranibizumab, and 30 eyes of combined treatment group underwent intravitreal injection of ranibizumab and macular grid photocoagulation 1wk later. The best corrected visual acuity ( BCVA ) , central macular thickness ( CMT ) measured by optical coherence tomography ( OCT ) and postoperative complications were observed.
RESULTS:ln simple injection group, the BCVA after operation were separately 0. 390 ± 0. 075 (4wk), 0. 367 ± 0. 088 (8wk) and 0. 319 ± 0. 064 (12wk), the CMT after operation were separately 221. 63 ± 112. 34μm (4wk), 337. 73±99. 56μm (8wk) and 432. 92 ± 100. 46μm (12wk), which were much better than pre-operation. But during follow-up, the BCVA presented down trend and the CMT was on the rise slowly. ln combined treatment group, the BCVA after operation were separately 0. 385 ± 0. 036 (4wk), 0.382±0.079 (8wk) and 0.377±0.097 (12wk),the CMT after operation were separately 249. 77 ± 106. 55μm (4wk), 270. 40 ± 92. 88μm (8wk) and 275. 84 ± 97. 34μm (12wk ), which were satisfactory and steady during follow-up, better than simple injection group (P<0. 05).CONCLUSlON:lntravitreal injection of ranibizumab can effectively improve visual acuity and decrease central foveal thickness for patients with DME, combining with macular grid photocoagulation can ensure therapeutic effects steady and permanent.

Objective To increase the transfection of EGFP-N3 plasmids into 293T cells using ultrasound-targeted microbubbles delivery(UTMD) mediated a peptide nucleic acid (PNA) binding nuclear localization signal (NLS).Methods Antibody-targeted microbubbles were used in the experiments which can specifically recognize the SV40T antigen receptor.The SV40T antigen receptors were expressed on the membranes of 293T cells.The PNA containing the NLS were inserted in the EGFP-N3 plasmid DNA,which increased nuclear localization.Ultrasound-targeted microbubble delivery (UTMD) and the PNA binding NLS were utilized to improve the cytoplasmic import of plasmids and the nuclear intake of the plasmid from the cytoplasm,respectively.The study was divided into five groups:Contrast (group A),Common microbubble + DNA (group B),Antibody-targeted microbubbles + DNA (group C),Common microbubbles + NLS-PNA-DNA (group D),Antibody-targeted microbubbles + NLS-PNA-DNA (group E).Fluorescence microscope was used to observe the fluorescent light in each group;flow cytometry to test the transfection;RT-PCR and Western blot to detect genes' mRNA and protein expression level.Results Ultrasound and antibody-targeted microbubble delivery (UTMD) significantly enhanced the cytoplasmic intake of exogenous genes and maintained high cell viability(＞80％).Fluorescent microscope showed that the quantities of green fluorescence in cells were increased successfully.The transfection results of flow cytometry were 0,(9.30 ± 0.46)％,(26.46 ± 2.01)％,(29.54 ± 0.62)％,(45.72 ± 1.86)％,respectively,and the differences were statistically significant(P ＜0.05).The relative mRNA and protein expression in group E were greater than those in group C and D respectively (P ＜0.05).Conclusions UTMD combined with antibody-targeted microbubbles and a PNA binding NLS plasmid can significantly improve transfection efficiency of exogenous genes by enhancing both cytoplasmic and nuclear DNA import.

The network teaching platform of ophthalmology of Qingdao University , as the basis construction of the national key discipline , forms a perfect and complete set of teaching system with the aid of part of course information, part of network teaching resources and part of answer and interaction. The network teaching platform focuses on the construction of network teaching resources and answer and interaction. In the college teaching of ophthalmology, through building clinical teaching resource database and discussing on network platform, we carry out interactive and discussion-based teaching, and students can prepare before class and discuss after class. The application of network teaching platform of ophthal-mology in classroom teaching and teaching feedback can improve the teaching methods , deepen the teach-ing content, implement the sharing of teaching resources, and lay a solid foundation for ophthalmology teaching reform.